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Evaluation of the effect of storage condition on cell extraction and flow cytometric analysis from intestinal

Manon E Wildenberg1, Marjolijn Duijvestein1, Liset Westera2

  • 1Tytgat Institute for Liver and Intestinal Research, Amsterdam, The Netherlands; Academic Medical Center, Department of Gastroenterology and Hepatology, Amsterdam, The Netherlands.

Journal of Immunological Methods
|May 18, 2018
PubMed

Insights

Storing intestinal biopsies at -20°C in DMSO/citrate buffer for up to 48 hours preserves viable immune cells for flow cytometry analysis. This method supports centralized analysis in clinical trials.

Area of Science:

  • Immunology
  • Gastroenterology
  • Biotechnology

Background:

  • Flow cytometry (FC) analysis of intestinal biopsies requires rapid processing for accurate immune cell phenotyping.
  • Cell viability and data integrity are compromised by delays in sample processing.

Purpose of the Study:

  • To evaluate the impact of various storage conditions on viable cell yield and immune cell proportions in intestinal biopsies.
  • To determine optimal storage protocols for maintaining sample quality for FC analysis.

Main Methods:

  • Intestinal biopsies (N=224) from Crohn's disease patients were stored under different conditions or processed immediately.
  • Cells were isolated, stained for immune markers, and analyzed by FC.

Main Results:

  • Storage at -80°C significantly decreased viable CD45+ cell counts.
  • Storage at -20°C in DMSO/citrate buffer yielded adequate cells for FC analysis.
  • CD3+ cell proportions decreased after 48h storage at 4°C in complete medium.

Conclusions:

  • Storage of intestinal biopsies at -20°C in DMSO/citrate buffer for up to 48 hours is suitable for FC analysis.
  • This storage method supports centralized FC analysis in multicenter clinical trials by enabling sample shipping and storage.
Abstract

Keywords:
Central analysisFlow cytometryInflammatory bowel diseaseMulticenter clinical trialTranslational research

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